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- NOTE] Bioluminescent Assay for Sphingolipid Ceramide N-Deacylase Using Vibrio harveyi Dark Mutant M-17
- Ki Woong Cho
- J. Microbiol. 2008;46(5):585-589. Published online October 31, 2008
- DOI: https://doi.org/10.1007/s12275-008-0114-2
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Abstract
PDF - A new bioluminescent assay method for the activity of sphingolipid ceramide N-deacylase (SCDase: EC3.5.1.69) as well as ceramidase (CDase: EC 3.5.1.23) was developed using bioluminescent marine bacteria. Enzymatically synthesized ceramide (N-myristoyl sphigosine, C14:0-18:1) and commercial SCDase were used in this demonstration, and myristic (tetradecanoic, C14:0) acid produced by the SCDase hydrolysis was quantified using Vibrio harveyi M-17, a dark mutant of V. harveyi. The in vivo light intensity of M-17 was stimulated up to thousands fold in the presence of myristic acid, was used for this assay. SCDase activity with as little as 10 μU and 5 nM of myristic acid production were detected in less than one min. <br>The assay worked well for the determination of Km and chromatographic fraction assay.
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Citations
Citations to this article as recorded by
- Control of metabolism and signaling of simple bioactive sphingolipids: Implications in disease
Patricia Gangoiti, Luz Camacho, Lide Arana, Alberto Ouro, Maria H. Granado, Leyre Brizuela, Josefina Casas, Gemma Fabriás, José Luis Abad, Antonio Delgado, Antonio Gómez-Muñoz
Progress in Lipid Research.2010; 49(4): 316. CrossRef
- Control of metabolism and signaling of simple bioactive sphingolipids: Implications in disease
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